Microparticle analyzing apparatus and data displaying method

ABSTRACT

Disclosed herein is a microparticle analyzing apparatus including a detecting portion configured to simultaneously detect a fluorescence generated from a microparticle in plural wavelength regions and a displaying portion configured to display thereon detection results in the plural wavelength regions in a form of a spectrum.

CROSS REFERENCES TO RELATED APPLICATIONS

The present application is a continuation of U.S. application Ser. No.16/184,616, filed Nov. 8, 2018, which is a continuation of U.S.application Ser. No. 15/460,493, filed on Mar. 16, 2017, now U.S. Pat.No. 10,147,209, issued on Dec. 4, 2018, which is a divisional of U.S.application Ser. No. 13/089,569, filed on Apr. 19, 2011, now U.S. Pat.No. 9,619,907, issued on Apr. 11, 2017, which claims priority toJapanese Priority Patent Application JP 2010-104520, filed in the JapanPatent Office on Apr. 28, 2010, the entire content of each of which ishereby incorporated by reference herein.

BACKGROUND

The present application relates to a microparticle analyzing apparatusand a method of displaying data obtained through a measurement by usingthe microparticle analyzing apparatus. More particularly, theapplication relates to a technique for detecting a light emitted from amicroparticle, and analyzing a kind of light thus emitted, and the like.

In general, when a biologically-relevant microparticle such as a cell, amicrobe or a liposome is analyzed, a flow cytometry (flow cytometer) isutilized. This technique, for example, is described in a non-patentliterary document of “Additional Volume of Cell Engineering ExperimentalProtocol Series Flow Cytometry Capable of being Manipulated withFreedom,” supervised by Takamitsu Nakauchi Shujunsha Co., Ltd. 2ndedition published on Aug. 31, 2006. The flow cytometry is based on amethod in which a laser beam (excited light) having a specificwavelength is radiated to microparticles which are caused to flow in aline within a flow path, and fluorescences or scattered lights emittedfrom the microparticles are detected, thereby analyzing the pluralmicroparticles one by one. With the flow cytometry, lights detected byrespective photodetectors are converted into electrical signals to bequantified, and a statistical analysis is carried out for the resultingelectrical signals thus quantified, thereby making it possible to judgekinds, sizes, structures, etc. of individual microparticles.

In addition, in recent years, in the flow cytometry, a multicoloranalysis using plural fluorescent dyes has been in widespread use. Thisflow cytometry, for example, is described in Japanese Patent Laid-OpenNo. 2006-230333 and JP-T-2008-500558. Each of the existing flowcytometries normally includes plural light sources corresponding todifferent wavelengths, respectively, and plural detectors for detectinglights emitted from respective dyes. On the other hand, since afluorescent dye has a spectrum, when plural fluorescent dyes are used inone measurement as with the multicolor analysis, lights from therespective fluorescent dyes other than an objective fluorescent dye areleaked to the detectors, thereby reducing the analysis precision. Inorder to cope with such a situation, with the existing flow cytometer,in order to extract only the optical information from the objectivefluorescent dye, mathematical correction, that is, fluorescencecorrection is carried out when the lights detected by using therespective photodetectors are converted into the electrical signals tobe quantified.

SUMMARY

As has been described, the fluorescent dyes have the spectra peculiarthereto, respectively, and the spectrum information thereof becomesimportant data representing the features of the fluorescent dyesthemselves. However, with the existing flow cytometer, the light fromthe objective fluorescent dye, for example, is detected by the sensorfor receiving a light having a specific bandwidth, and the data detectedby the sensor is treated as corresponding data. Therefore, there iscaused a problem that it may be difficult to propose the spectruminformation on the fluorescent dyes.

The present application has been made in order to solve the problemsdescribed above, and it is therefore primarily desirable to provide amicroparticle analyzing apparatus and a data displaying method each ofwhich is capable of measuring spectra of fluorescences generated frommicroparticles, respectively, and visually understanding measured databy a user.

In order to attain the desire described above, according to anembodiment, there is provided a microparticle analyzing apparatusincluding: a detecting portion configured to simultaneously detect afluorescence generated from a microparticle in plural wavelengthregions; and a displaying portion configured to display thereondetection results in the plural wavelength regions in a form of aspectrum.

According to the embodiment, since it is possible to propose thespectrum information on the fluorescence generated from themicroparticle, the user can visually understand the measured datairrespective of presence or absence of the fluorescence correction, andthe results of the fluorescence correction. Also, for example, the dataon the specific region is extracted, thereby obtaining the various kindsof pieces of information on the microparticle.

According to another embodiment, there is provided a data displayingmethod including the steps of: simultaneously detecting a fluorescencegenerated from a microparticle in plural wavelength regions; anddisplaying detected data for each wavelength region in a form of aspectrum.

As set forth hereinabove, according to the present application, since itis possible to propose the spectrum information on the fluorescencegenerated from the microparticle, the user can visually understand themeasured data.

Additional features and advantages are described herein, and will beapparent from the following Detailed Description and the figures.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a block diagram showing a configuration of a microparticleanalyzing apparatus according to a second embodiment;

FIG. 2 is a view showing an example of display of measurement results;

FIG. 3 is a view showing a method of applying gate to the example of thedisplay of the measurement results shown in FIG. 2; and

FIG. 4 is a view showing an example of display obtained after completionof the application of the gate.

DETAILED DESCRIPTION

Embodiments of the present application will be described below in detailwith reference to the drawings.

It is noted that the present application is by no means limited toembodiments which will be described below. In addition, the descriptionwill now be given in accordance with the following order.

1. First Embodiment (A method of displaying a spectrum)

2. Second Embodiment (A microparticle analyzing apparatus for displayinga spectrum)

1. First Embodiment

Entire Constitution

Firstly, a data displaying method according to a first embodiment willbe described. With the data displaying method according to the firstembodiment, a fluorescence generated from a microparticle such as a cellor a microbead is simultaneously detected in plural wavelength regions,and detected data for each wavelength region is displayed in the form ofa spectrum.

Detected Data

With the data displaying method of the first embodiment, a lightdetected by a photodetector is converted into a voltage pulse(electrical signal) every wavelength region, and the resulting voltagepulse is quantified to obtain detected data. Specifically, a height, awidth and an area of the voltage pulse are obtained, thereby obtainedthe detected data.

Display Form

With the data displaying method of the first embodiment, the detecteddata, for each wavelength region, obtained by using the method describedabove is displayed in the form of a spectrum. Although the datadisplaying method is especially by no means limited, for example, adensity spectrum, a dot plot, a contour line or the like is given. Inaddition, it is possible that only data in a specific region isextracted from the spectrum being displayed, an image of a spectrumchart is enlarged, a histogram or a two-dimensional plot is displayed,and so forth.

In addition, with the data displaying method of the first embodiment, itis also possible that the detected data which is acquired bycontinuously detecting plural microparticles is arithmetically operated,and results of the arithmetical operation are displayed. With regard tothe arithmetic operation, for example, there are given an accumulationof the detected data, a calculation of an average value of the detecteddata on all the microparticles, and the like. Moreover, with the datadisplaying method of the first embodiment, although the display can becarried out while the detection is carried out, the detected data may betemporarily preserved, and the data thus preserved may be read out anddisplayed. In this case as well, the detected data may be arithmeticallyoperated and results of the arithmetical operation may be displayed.

With the existing two-dimensional plot, even when the fluorescencecorrection is carried out, it is difficult for a user to visually judgethe data obtained through the fluorescence correction. On the otherhand, with the data displaying method of the first embodiment, since itis possible to propose the spectrum information on the fluorescencegenerated from the microparticle, the user can visually understand themeasured data irrespective of presence or absence of the fluorescencecorrection, and the results of the fluorescence correction. Also, withthe data displaying method of the first embodiment, the data in thespecific region is extracted, thereby making it possible to obtain thevarious kinds of pieces of information on the microparticle.

The data displaying method of the first embodiment is by no meanslimited to application to an analyzing apparatus for the microparticle,the cell or the like. For example, the data displaying method of thefirst embodiment can also be applied to manufacturing equipment,inspection equipment, medical equipment, or the like.

2. Second Embodiment

Entire Configuration of Apparatus

Next, a microparticle analyzing apparatus according to a secondembodiment will be described. FIG. 1 is a block diagram showing aconfiguration of the microparticle analyzing apparatus according to thesecond embodiment. Also, FIG. 2 is a view showing an example of displayof measurement results obtained from the microparticle analyzingapparatus of the second embodiment. As shown in FIG. 1, themicroparticle analyzing apparatus 1 is provided with at least adetecting portion 2, a converting portion 3, an analyzing portion 4, anda displaying portion 5.

Configuration of Detecting Portion 2

All it takes is that the detecting portion 2 has a configuration withwhich a fluorescence generated from a microparticle as an object of ananalysis can be simultaneously detected in plural wavelength regions.Specifically, the detecting portion 2 can have a configuration in whichsensors capable of detecting wavelength regions are disposed so as tocorrespond to the wavelength regions, respectively, or a configurationincluding a detector capable of simultaneously detecting plural lightsas with a multichannel Photo-Multiplier Tube (PMT) or the like. All ittakes is that the number of wavelength regions detected by the detectingportion 2, that is, the number of channels of the detector disposed inthe detecting portion 2 or the number of sensors disposed in thedetecting portion 2 is equal to or larger than the number of dyes used.In this case, preferably, the number of channels of the detectordisposed in the detecting portion 2 or the number of sensors disposed inthe detecting portion 2 is equal to or larger than twelve because in thecurrent multicolor analysis, six to eight colors are general, and ten totwelve colors are used at the most.

In addition, the microparticle analyzing apparatus 1 of the secondembodiment may also adopt such a construction that a spectroscope isprovided in the detector 2, and after the fluorescence generated fromthe microparticle is spectrally diffracted in the spectroscope, thefluorescence thus spectrally diffracted is made incident to the detectorsuch as the multichannel PMT. Moreover, an objective lens, a condensinglens, a pin hole, a band-cut filter, a dichroic mirror, or the like canalso be provided as may be necessary.

Configuration of Converting Portion 3

The converting portion 3 operates to convert the lights, in therespective wavelength regions, detected by the detecting portion 2 intovoltage pulses (electrical signals), respectively. For example, an ADconverter or the like can be used as the converting portion 3.

Configuration of Analyzing Portion 4

With the analyzing portion 4, the voltage pulses obtained through the ADconversion in the converting portion 3 are processed by using anelectronic computer or the like to quantify the voltage pulses, therebyobtaining the detected data. Specifically, a height (peak), a width oran area (integral) of the voltage pulse is obtained, and is used as thedetected data. Also, the results are associated with the wavelengthregions, respectively, and are preserved in a memory portion (not shown)or the like.

Configuration of Displaying Portion 5

The displaying portion 5 operates to display thereon the detected dataobtained in the processing in the analyzing portion 4, that is, themeasurement results obtained from the microparticle analyzing apparatus1 of the second embodiment. Specifically, for example, as shown in FIG.2, “a density plot” representing a fluorescence intensity for eachchannel, or the like is displayed on the displaying portion 5. In FIG.2, an axis of abscissa represents a channel number of the detectingportion 2, and an axis of ordinate represents an intensity. When thedetected wavelength is made short as the channel number becomes larger,the spectrum information on the detected light (fluorescence) isobtained.

Operation of Microparticle Analyzing Apparatus 1

Next, an operation of the microparticle analyzing apparatus 1 of thesecond embodiment will be described. Although the microparticle analyzedby using the microparticle analyzing apparatus 1 is especially by nomeans limited, for example, the cell, the microbead or the like isgiven. In addition, the kinds and the number of fluorescent dyesmodifying such microparticles are also especially by no means limited.Thus, the known dyes such as fluorescein isothiocyanate (C₂₁H₁₁NO₅S:FITC), phycoerythrin (PE), peridinin chlorophyll protein (PerCP), PE-Cy5and PE-Cy7 can be suitably selected and used as may be necessary. Inaddition, each of the microparticles may be modified with pluralfluorescent dyes.

When the microparticle is intended to be optically analyzed by using themicroparticle analyzing apparatus 1, firstly, an excited light isemitted from a light source of a light radiating portion to be radiatedto the microparticle which is being caused to pass within the flow path.Secondary, a fluorescence generated from the microparticle is detectedin the detecting portion 2. In this case, for example, the fluorescencegenerated from the microparticle is spectrally diffracted in aspectroscope such as a prism or a diffraction grating, whereby theresulting lights having different wavelength regions, respectively, aremade incident to the sensors or the channels of the PMT disposed in thedetecting portion 2.

After that, the data, in the wavelength regions, acquired in thedetecting portion 2 is converted into voltage pulses in the convertingportion 3, and the resulting voltage pulses are quantified in theanalyzing portion 4 to be preserved therein. The results of theanalysis, for example, are displayed in the form of “the density plot”shown in FIG. 2, or the like on the displaying portion 5. It is notedthat “the density plot” shown in FIG. 2 is the results obtained bymeasuring a sample obtained by mixing a sample modified with only FITC,a sample modified with only PE, and a Negative sample with one anotherby using a 32-channel PMT.

For “the density plot,” a channel number (wavelength-dependent number)of the PMT is plotted on an axis of abscissa, and a fluorescenceintensity is plotted on an axis of ordinate. In this case, the number,0, of accumulation is set as blue, and red comes to be obtained as thenumber of accumulation is increased, that is, the density is increased.At this time, when the number of the channel corresponding to theshortest detected wavelength is set as 1, and the detected wavelengthbecomes short as the number of the channel becomes larger, informationcorresponding to the fluorescence spectrum is obtained. Also, from thespectrum information, the user can recognize what kind of fluorescencethe sample is labeled with.

In addition, with the microparticle analyzing apparatus 1 of the secondembodiment, it is also possible to carry out the gating for selectingonly the specific sample group with respect to the data shown in FIG. 2,and extracting and displaying the data on only the specific sample group(microparticles). The display form in this case is especially by nomeans limited, and thus the data concerned can be displayed in the formof various kinds of forms such as a histogram, a two-dimensional plotand a spectrum chart. FIG. 3 is a view showing a method of applying thegate to the example of the display shown in FIG. 2, and FIG. 4 is a viewshowing an example of display after completion of the application of thegate. Since in the existing flow cytometer, normally, the gatingprocessing is executed for the two-dimensional plot, the gate is appliedto only the information for two channels.

On the other hand, as shown in FIG. 3, in the microparticle analyzingapparatus 1 of the second embodiment, the gate, for example, is appliedwith ten channels as a batch with respect to the fluorescence spectrum.Also, as shown in FIG. 4, for the data on the sample to which the gatingis applied, a dot-plot mode can be adopted in the spectrum plot, or acolor can be changed in the spectrum plot. As a result, it is possibleto realize the more explicit gating, and thus the user can clearlyconfirm the data concerned. In addition, the data on the sample to whichthe gating is applied can also be displayed in the form of atwo-dimensional plot.

In addition, the region to which the gating is to be applied can beselected based on the fluorescence spectrum. Therefore, for example,whether or not two kinds of fluorescent dyes are fixed to onemicroparticle, or whether or not two microparticles modified withdifferent fluorescent dyes, respectively, are contained can be easilysheared. In addition, the gating is applied to a portion to which nofluorescence is leaked from any other fluorescent dyes, whereby the usercan visually get the various kinds of pieces of information from thedata being displayed irrespective of presence or absence of thefluorescence correction or the results of the fluorescence correction.

It should be noted that with the microparticle analyzing apparatus 1 ofthe second embodiment, not only “the density plot” or “the dot plot”described above, but also “a contour line plot,” “a two-dimensionalplot,” “a two-dimensional histogram” or the like can be displayed. Inaddition, the characteristics of plural microparticles can becontinuously measured, and the data detected in the detecting portion 2is accumulated at any time and can be reflected in the fluorescencespectrum being displayed on the displaying portion 5 at real time.Moreover, the data preserved can be read out and displayed, and theresults of continuously measuring the characteristics of pluralmicroparticles can be temporarily preserved and can be displayed byusing an average value of all the samples.

It should be understood that various changes and modifications to thepresently preferred embodiments described herein will be apparent tothose skilled in the art. Such changes and modifications can be madewithout departing from the spirit and scope and without diminishing itsintended advantages. It is therefore intended that such changes andmodifications be covered by the appended claims.

The application is claimed as follows:
 1. A system comprising: a lightsource for irradiating light to each of microparticles stained with aplurality of fluorescent dyes; a plurality of detectors configured todetect a fluorescence generated from the each of microparticles inplural wavelength regions; and a display configured to display thereon avisual representation, wherein the visual representation, representingdetected data in each one of the plural wavelength regions, has aplurality of colors, and a respective color at each fluorescenceintensity in each wavelength region indicates a number of microparticlesat said fluorescence intensity in said wavelength region, wherein eachdetector corresponds to a wavelength region, and wherein a number of thefluorescent dyes is equal or less than a number of the detectors.
 2. Thesystem of claim 1, wherein the light source and the plurality ofdetectors are included in a flow cytometry apparatus.
 3. The system ofclaim 1, further comprising a flow path for passing the plurality ofmicroparticles stained with the plurality of fluorescent dyes.
 4. Thesystem of claim 1, further comprising a memory configured to store thedetected data.
 5. The system of claim 1, further comprising a secondlight source for irradiating light to the each of microparticles passingthrough the flow path.
 6. The system of claim 1, further comprising aprism.
 7. The system of claim 1, further comprising diffraction grating.8. The system of claim 1, wherein fluorescence generated from the onemicroparticle is spectrally diffracted in a spectroscope.
 9. The systemof claim 1, further comprising an AD converter that converts thefluorescence detected by the detectors into electrical signals, whereinthe processer configured to generate the detected data from theelectrical signals.
 10. The system of claim 4, wherein the storeddetected data is read out and displayed.
 11. The system of claim 1,wherein the number of the fluorescent dyes is zero or one.
 12. Thesystem of claim 1, wherein the number of the fluorescent dyes is lessthan twelve.
 13. The system of claim 1, wherein the microparticles arecells.
 14. The system of claim 1, wherein the fluorescent dyes includeat least one of FITC, PE, PerCP, PE-Cy5, and PE-Cy7.
 15. The system ofclaim 1, wherein the visual representation is at least one of a densityplot, a dot plot and a contour line plot.
 16. The system of claim 1,wherein the detected data is mapped for each one of the pluralwavelength regions and each fluorescence intensity in the visualrepresentation.
 17. The system of claim 1, wherein each of the pluralwavelength regions is displayed with a respective channel number. 18.The system of claim 1, wherein only data in a specific region selectedfrom the visual representation is extracted and displayed.
 19. Thesystem of claim 18, wherein the data extracted is two-dimensionallyplotted.
 20. The system of claim 1, wherein the detected data includesat least one of height, width and area of the electrical signal.
 21. Thesystem of claim 1, wherein the wavelength regions include a firstwavelength region of a first width and a second wavelength region of asecond width.
 22. The system of claim 1, wherein the plural wavelengthregions include twelve different wavelength regions.
 23. The system ofclaim 1, wherein the plural wavelength regions include more than twelvedifferent wavelength regions.
 24. A flow cytometry method comprising:passing, through a flow path in a flow cytometry apparatus, a pluralityof microparticles stained with a plurality of fluorescent dyes;irradiating light, from a light source, to each of the microparticlespassing through the flow path; detecting, by a plurality of detectors, afluorescence generated from the each of the microparticles in pluralwavelength regions; and displaying a visual representation on a display,wherein the visual representation, representing detected data in eachone of the plural wavelength regions, has a plurality of colors, and arespective color at each fluorescence intensity in each wavelengthregion indicates a number of microparticles at said fluorescenceintensity in said wavelength region, wherein each detector correspondsto a wavelength region, and wherein a number of the fluorescent dyes isequal or less than a number of the detectors.
 25. The method of claim24, wherein the number of the fluorescent dyes is zero or one.
 26. Themethod of claim 24, wherein the number of the fluorescent dyes is lessthan twelve.
 27. The method of claim 24, wherein the microparticles arecells.
 28. The method of claim 24, wherein the fluorescent dyes includeat least one of FITC, PE, PerCP, PE-Cy5, and PE-Cy7.
 29. The method ofclaim 24, wherein the visual representation is at least one of a densityplot, a dot plot and a contour line plot.
 30. The method of claim 24,wherein the detected data is mapped for each one of the pluralwavelength regions and each fluorescence intensity in the visualrepresentation.
 31. The method of claim 24, wherein each of the pluralwavelength regions is displayed with a respective channel number. 32.The method of claim 24, wherein only data in a specific region selectedfrom the visual representation is extracted and displayed.
 33. Themethod of claim 32, wherein the data extracted is two-dimensionallyplotted.
 34. The method of claim 24, wherein the detected data includesat least one of height, width and area of the electrical signal.
 35. Themethod of claim 24, wherein the wavelength regions include a firstwavelength region of a first width and a second wavelength region of asecond width.
 36. The method of claim 24, wherein the plural wavelengthregions include twelve different wavelength regions.
 37. The method ofclaim 24, wherein the plural wavelength regions include more than twelvedifferent wavelength regions.
 38. A flow cytometry apparatus comprising:a flow path for passing a plurality of microparticles stained with aplurality of fluorescent dyes; a light source for irradiating light toeach of the microparticles passing through the flow path; a plurality ofdetectors configured to detect a fluorescence generated from the each ofthe microparticles in plural wavelength regions; and a processorcircuitry configured to generate detected data related to thefluorescence and output the detected data for displaying a visualrepresentation on a display, wherein the visual representation,representing the detected data in each one of the plural wavelengthregions, has a plurality of colors, and a respective color at eachfluorescence intensity in each wavelength region indicates a number ofmicroparticles at said fluorescence intensity in said wavelength region,wherein each detector corresponds to a wavelength region, and wherein anumber of the fluorescent dyes is equal or less than a number of thedetectors.
 39. The flow cytometry apparatus of claim 38, furthercomprising a second light source for irradiating light to the each ofmicroparticles passing through the flow path.
 40. The flow cytometryapparatus of claim 38, further comprising a prism.
 41. The flowcytometry apparatus of claim 38, further comprising diffraction grating.42. The flow cytometry apparatus of claim 38, wherein fluorescencegenerated from the one microparticle is spectrally diffracted in aspectroscope.
 43. The flow cytometry apparatus of claim 38, furthercomprising an AD converter that converts the fluorescence detected bythe detectors into electrical signals, wherein the processer configuredto generate the detected data from the electrical signals.
 44. The flowcytometry apparatus of claim 38, wherein the number of the fluorescentdyes is zero or one.
 45. The flow cytometry apparatus of claim 38,wherein the number of the fluorescent dyes is less than twelve.
 46. Theflow cytometry apparatus of claim 38, wherein the microparticles arecells.
 47. The flow cytometry apparatus of claim 38, wherein thefluorescent dyes include at least one of FITC, PE, PerCP, PE-Cy5, andPE-Cy7.
 48. The flow cytometry apparatus of claim 38, wherein the visualrepresentation is at least one of a density plot, a dot plot and acontour line plot.
 49. The flow cytometry apparatus of claim 38, whereinthe detected data is mapped for each one of the plural wavelengthregions and each fluorescence intensity in the visual representation.50. The flow cytometry apparatus of claim 38, wherein each of the pluralwavelength regions is displayed with a respective channel number. 51.The flow cytometry apparatus of claim 38, wherein only data in aspecific region selected from the visual representation is extracted anddisplayed.
 52. The flow cytometry apparatus of claim 51, wherein thedata extracted is two-dimensionally plotted.
 53. The flow cytometryapparatus of claim 38, wherein the detected data includes at least oneof height, width and area of the electrical signal.
 54. The flowcytometry apparatus of claim 38, wherein the wavelength regions includea first wavelength region of a first width and a second wavelengthregion of a second width.
 55. The flow cytometry apparatus of claim 38,wherein the plural wavelength regions include twelve differentwavelength regions.
 56. The flow cytometry apparatus of claim 38,wherein the plural wavelength regions include more than twelve differentwavelength regions.
 57. A non-transitory computer readable storagemedium storing one or more computer programs adapted to cause aprocessor based system to execute steps comprising: receiving detecteddata related to the fluorescence detected from each of microparticlesstained with a plurality of fluorescence dyes in plural wavelengthregions by a plurality of detectors; and displaying a visualrepresentation on a display, wherein the visual representation,representing the detected data in each one of the plural wavelengthregions, has a plurality of colors, and a respective color at eachfluorescence intensity in each wavelength region indicates a number ofmicroparticles at said fluorescence intensity in said wavelength region,wherein each detector corresponds to a wavelength region, and wherein anumber of the fluorescent dyes is equal or less than a number of thedetectors.
 58. The non-transitory computer readable storage medium ofclaim 57, wherein the plurality of detectors are included in a flowcytometry apparatus.
 59. The non-transitory computer readable storagemedium of claim 57, wherein the number of the fluorescent dyes is zeroor one.
 60. The non-transitory computer readable storage medium of claim57, wherein the number of the fluorescent dyes is less than twelve. 61.The non-transitory computer readable storage medium of claim 57, whereinthe microparticles are cells.
 62. The non-transitory computer readablestorage medium of claim 57, wherein the fluorescent dyes include atleast one of FITC, PE, PerCP, PE-Cy5, and PE-Cy7.
 63. The non-transitorycomputer readable storage medium of claim 57, wherein the visualrepresentation is at least one of a density plot, a dot plot and acontour line plot.
 64. The non-transitory computer readable storagemedium of claim 57, wherein the detected data is mapped for each one ofthe plural wavelength regions and each fluorescence intensity in thevisual representation.
 65. The non-transitory computer readable storagemedium of claim 57, wherein each of the plural wavelength regions isdisplayed with a respective channel number.
 66. The non-transitorycomputer readable storage medium of claim 57, wherein only data in aspecific region selected from the visual representation is extracted anddisplayed.
 67. The non-transitory computer readable storage medium ofclaim 66, wherein the data extracted is two-dimensionally plotted. 68.The non-transitory computer readable storage medium of claim 57, whereinthe detected data includes at least one of height, width and area of theelectrical signal.
 69. The non-transitory computer readable storagemedium of claim 57, wherein the wavelength regions include a firstwavelength region of a first width and a second wavelength region of asecond width.
 70. The non-transitory computer readable storage medium ofclaim 57, wherein the plural wavelength regions include twelve differentwavelength regions.
 71. The non-transitory computer readable storagemedium of claim 57, wherein the plural wavelength regions include morethan twelve different wavelength regions.